title: CIN85 modulates the down-regulation of Fc gammaRIIa expression and function by c-Cbl in a PKC-dependent manner in human neutrophils.
abstract: We previously described a non-classical mechanism that arrests FcγRIIa signaling in human neutrophils once engaged by immune complexes or opsonized pathogens. The engagement of FcγRIIa leads to its ubiquitination by the ubiquitin ligase c-Cbl and degradation by the proteasome. Herein, we further examined some of the events regulating this novel pathway. The adaptor protein CIN85 was described in other systems to be involved in the regulation of the c-Cbl-dependent pathway. We found that CIN85 is expressed in human neutrophils and that it translocates like c-Cbl from the cytosol to the plasma membrane following receptor cross-linking. CIN85 was also recruited to the same subset of high density detergent-resistant membrane fractions in which stimulated FcγRIIa partitioned with c-Cbl. The integrity of these microdomains is essential to the FcγRIIa degradation process because the cholesterol-depleting agent methyl-β-cyclodextrin inhibits this event. Silencing the expression of CIN85 by siRNA in dibutyryl cyclic AMP-differentiated PLB 985 cells prevented FcγRIIa degradation and increased IgG-mediated phagocytosis. Confocal microscopy revealed that the presence of CIN85 is essential to the proper sorting of FcγRIIa during endocytosis. We also provide direct evidence that CIN85 is a substrate of serine/threonine kinase PKCs. Classical PKCs positively regulate FcγRIIa ubiquitination and degradation because these events were inhibited by Gö6976, a classical PKC inhibitor. We conclude that the ubiquitination and degradation of stimulated FcγRIIa mediated by c-Cbl are positively regulated by the adaptor protein CIN85 in a PKC-dependent manner and that these events contribute to the termination of FcγRIIa signaling.
authors.affiliation: Centre de Recherche du CHUQ, Département de Microbiologie-Infectiologie et Immunologie, Faculté de Médecine, Université Laval, Québec, Québec G1V 4G2, Canada.
authors.forename: Louis
authors.initials: L
authors.lastname: Marois
authors.forename: Myriam
authors.initials: M
authors.lastname: Vaillancourt
authors.forename: Guillaume
authors.initials: G
authors.lastname: Paré
authors.forename: Valérie
authors.initials: V
authors.lastname: Gagné
authors.forename: Maria J G
authors.initials: MJ
authors.lastname: Fernandes
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Rollet-Labelle
authors.forename: Paul H
authors.initials: PH
authors.lastname: Naccache
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism / Q000502:physiology*; D015536:Down-Regulation / Q000235:genetics; D006801:Humans; D009504:Neutrophils / Q000378:metabolism*; D011493:Protein Kinase C / Q000378:metabolism*; D055550:Protein Stability; D021381:Protein Transport; D050721:Proto-Oncogene Proteins c-cbl / Q000502:physiology*; D017452:Receptors, IgG / Q000096:biosynthesis*; D015398:Signal Transduction / Q000276:immunology; D054875:Ubiquitination
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C080568:Fc gamma receptor IIA; D017452:Receptors, IgG; C433810:SH3KBP1 protein, human; D050721:Proto-Oncogene Proteins c-cbl; D011493:Protein Kinase C; C067419:CBL protein, human